Glycerophosphoinositol in preventing and treating covid-19 infections and method for obtaining it

Abstract

Glycerophosphoinositol (GPI) is used in preventing and treating COVID-19 infections. An environmentally sustainable method is for obtaining glycerophosphoinositol. In particular, a process for preparing glycerophosphoinositol from crude or partially purified phospholipid mixtures, includes in sequence: a) hydrolysis of a crude or partially purified phospholipid mixture by treatment with PLA1 and PLA2 enzymes; b) microfiltration of the mixture from step a) and subsequent ultrafiltration and nanofiltration of the microfiltrate to give a concentrated aqueous fraction of reaction products; c) electrodialysis of the aqueous fraction of step b) for separating ionic compounds from neutral compounds; d) ion exchange chromatography. Moreover, glycerophosphoinositol is used in preventing and treating a COVID-19 syndrome.

1 . A process for preparing glycerophosphoinositol from crude or partially purified phospholipid mixtures, comprising the following steps in sequence:
 a) hydrolysis of a crude or partially purified phospholipid mixture by treatment with PLA1 and PLA2 enzymes;   b) microfiltration of the mixture from step a) to obtain a microfiltrate, and subsequent ultrafiltration and nanofiltration of the microfiltrate to give a concentrated aqueous fraction of reaction products;   c) electrodialysis of the aqueous fraction of step b) for separating ionic compounds from neutral compounds;   d) ion exchange chromatography.   
     
     
         2 . The process according to  claim 1 , wherein the crude or partially purified phospholipid mixture of step a) has a phospholipid concentration in water between 10 and 100 g/kg of mixture. 
     
     
         3 . The process according to  claim 2 , wherein the phospholipid mixture is a fine dispersion of lecithin in water, wherein lecithin has a concentration between 45 and 55 g/kg. 
     
     
         4 . The process according to  claim 1 , wherein an amount of PLA1 and PLA2 enzyme in step a) is in a range between 200 and 4000 mg/kg of mixture. 
     
     
         5 . The process according to  claim 4 , wherein step a) is performed with a mixture of PLA1 and PLA2, wherein a concentration of PLA1 and a concentration of PLA2 are between 800 and 1300 mg/kg of aqueous suspension. 
     
     
         6 . The process according to  claim 1 , wherein step a) is performed at a pH between 3.5 and 5.5 and at a temperature between 15° C. and 55° C., and wherein an initial pH of the reaction mixture of about 6.5 is brought to a range of pH=3.5-5 by addition of concentrated phosphoric acid. 
     
     
         7 . The process according to  claim 1 , wherein in step b) the pH is adjusted to about 7 by addition of a base comprising a concentrated NaOH or KOH solution. 
     
     
         8 . The process according to  claim 1 , wherein in step b) the microfiltration is performed by tangential filtration on polymer membranes, ceramic membranes, or steel membranes having a porosity between 0.45 and 0.1 micron, and wherein the microfiltration is followed in sequence by a diafiltration. 
     
     
         9 . The process according to  claim 1 , comprising a first ultrafiltration and, a second ultrafiltration, wherein the first ultrafiltration and the second ultrafiltration are performed on wound spiral polymer membranes, and wherein:
 the first ultrafiltration is made using membranes with a cut-off in the range from 20 kDa to 500 kDa,   the second ultrafiltration is made using membranes with a cut-off in the range from 1 kDa to 10 kDa,   and wherein the nanofiltration is performed on wound spiral polymer membranes, with a cut-off value ranging from 150 Da to 3000 Da.   
     
     
         10 . The process according to  claim 9 , wherein step b) comprises the following sub-steps:
 b1) ultrafiltration of the microfiltration product with a wound spiral polymer PES membrane with a cut-off value of about 50,000 Da;   b2) subsequent ultrafiltration with a membrane having a cut-off of about 5000 Da, wherein GPI, GPE, GPC and other molecules with low molecular weight permeate and further oligomers recirculate;   b3) membrane nanofiltration with a cut-off value of about 1000-150 Da, with a wound spiral PES membrane, to obtain a partial separation between GPI, kept under recirculation, and GPC and GPE passing with the permeate;   b4) diafiltration with demineralized water with low conductivity to obtain a recirculation solution mainly containing GPI and relatively lower concentrations of GPC and GPE with respect to the fraction of step b3).   
     
     
         11 . The process according to  claim 1 , wherein in step c) an aqueous solution is circulated through an electrodialysis system, to obtain migration of the GPI salts and the preservation of GPC and GPE in a main flow solution. 
     
     
         12 . The process according to  claim 1 , wherein the permeated solution is further subjected to a second step of nanofiltration with membranes in the range from 1000 Da to 150 Da to eliminate mainly inorganic substances with low molecular weight and to concentrate the solution. 
     
     
         13 . The process according to  claim 1 , wherein step d) of ion exchange chromatography comprises i) preliminary ion exchange on strong cationic resin in H+ form of the aqueous solution containing GPI, GPE and GPC at varying concentrations, and then ii) passage of the eluted solution on anionic resin in strong or weak OH— form, and wherein, if using weak anionic resins, the resin is first filled with solution from the acid exchange, then washed with water, with a diluted solution of formic, acetic or propionic acid at a concentration of about 1-3% w/v to remove less acidic impurities, and then eluted with a solution of formic, acetic or propionic acid at 5-10% w/v to obtain a pure GPI solution. 
     
     
         14 . The process according to  claim 13 , wherein step d) is performed with a strong sulfonic resin in H+ form, followed by a weak resin in OH— form, wherein the weak base resin is first washed with 12 g/l acetic acid and then eluted with ammonium carbonate at a concentration of 0.03 M. 
     
     
         15 - 18 . (canceled) 
     
     
         19 . A method for preventing or early treating a syndrome due to COVID-19 or other viral infections, comprising administering an effective amount of glycerophosphoinositol purified as obtained by the process according to  claim 1 . 
     
     
         20 . The method according to claim  15 , wherein the syndrome is sepsis due to COVID-19 or other viral infections.